pET-31b(+)

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pET31b载体基本信息

出品公司: EMD Biosciences (Novagen)
别名: pET31b, pet-31b(+)
质粒类型: 大肠杆菌蛋白表达
表达水平:
克隆方法: 多克隆位点,限制性内切酶
载体大小: 5742 bp
表达宿主菌: BL21(DE3) Rosetta2(DE3)C43(DE3)
5' 测序引物: T7: 5'-TAATACGACTCACTATAGGG-3'
3' 测序引物: T7t: 5'-GCTAGTTATTGCTCAGCGG-3'
载体标签: C-His, N-KSI
载体抗性: 氨苄
备注: 融合表达有N端酮类固醇异构酶,有助于提高蛋白及小肽的产量; AlwNI单克隆位点可以用来插入单向重复多肽序列。
产品目录号: 69952-3
稳定性: 瞬时表达
组成型: 组成型
病毒/非病毒: 非病毒

pET31b载体质粒图谱和多克隆位点信息

pet31b载体图谱
pet31b多克隆位点

pET31b载体简介

pET-31b(+)载体(Cat. No. 69952-3)是设计用来高水平表达蛋白质的载体,载体融合表达有125个氨基酸的酮类固醇异构酶(ketosteroid isomerase protein) 。载体上面的AlwN I单克隆位点允许单向插入串联重复的通过蛋氨酸(methionine)分隔开的多肽序列。

pET31b载体的单一的多克隆位点见上面的环状质粒图谱。注意:载体序列是以pBR322质粒的编码规矩进行编码的,所以T7蛋白表达区在质粒图谱上面是反向的。

T7 RNA聚合酶启动的克隆和表达区域在质粒图谱中也被标注了出来。质粒的F1复制子是被定向的,所以在T7噬菌体聚合酶的作用下,包含有蛋白编码序列的病毒 粒子能够产生,并启动蛋白表达,同时蛋白表达将被T7终止子序列(Cat. No. 69337-3)的作用下终止蛋白翻译。

pET31b载体序列

LOCUS       pET-31b(+)    5742 bp     DNA  Circular    SYN
DEFINITION  pET-31b(+)
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTOAUTHORAD1|Novagen Inc.|
FEATURES             Location/Qualifiers
     source          1..5742
                     /organism="pET-31b(+)"
                     /mol_type="other DNA"
     terminator      complement(1..129)
                     /label="T7_terminator"
     misc_feature    69..87
                     /label="T7_Terminal_primer"
     misc_feature    complement(553..569)
                     /label="T7_transl_en_RBS"
     misc_feature    complement(587..614)
                     /label="lacO"
     promoter        complement(614..632)
                     /label="T7_promoter"
     gene            664..927
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
     misc_feature    complement(716..735)
                     /label="pBRrevBam_primer"
     CDS             769..1269
                     /label="ORF frame 1"
     misc_feature    1010..2101
                     /label="lacI"
     CDS             1142..2101
                     /label="ORF frame 2"
     gene            2160..2435
                     /label="tet (576 - 851)"
                     /gene="tet (576 - 851)"
     misc_feature    2910..3101
                     /label="ROP"
     misc_feature    complement(3095..3117)
                     /label="pGEX_3_primer"
     gene            complement(4284..5144)
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             complement(4284..5144)
                     /label="ORF frame 2"
     promoter        complement(5186..5214)
                     /label="AmpR_promoter"
ORIGIN
    1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
   61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
  121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagcagcatc tggcatgcgt
  181 gaatattctt ctcgccaaac aaggcgcgga tgctcaccac cttgccggcg ccattgaagc
  241 gaaagtgatc gatgggcgca actacggtct tgcggccctg atactcgaag ctgacggtga
  301 aagcgaaggc cgcttcgttg gcgaccgcgc gtacctcctg cgtcagctcc accgccaaag
  361 gcagtttgag cgagttggcg taaaactcac gaatcgcagc cgtaccggac ctgggctcgg
  421 aacccacggg gtcttccacc gtggcgtcat cggcaaacag cgcgacgatg ccgtccagat
  481 cgccggcatt gagcgcagcc acaaagcgct gtaccacggc ggtgatgtgt tctggggtat
  541 gcatatgtat atctccttct taaagttaaa caaaattatt tctagagggg aattgttatc
  601 cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc
  661 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat
  721 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
  781 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg
  841 catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc
  901 ctaatgcagg agtcgcataa gggagagcgt cgagatcccg gacaccatcg aatggcgcaa
  961 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt
 1021 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
 1081 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
 1141 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
 1201 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
 1261 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
 1321 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
 1381 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
 1441 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
 1501 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
 1561 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
 1621 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
 1681 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
 1741 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
 1801 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
 1861 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
 1921 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
 1981 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
 2041 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
 2101 agcgcaacgc aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc
 2161 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
 2221 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
 2281 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
 2341 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
 2401 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cccacgggtg cgcatgatcg
 2461 tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg
 2521 aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag
 2581 caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag
 2641 cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa
 2701 cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc
 2761 gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat
 2821 catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga
 2881 acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca
 2941 tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg
 3001 cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca
 3061 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
 3121 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
 3181 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
 3241 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatatgcgg
 3301 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
 3361 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
 3421 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
 3481 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
 3541 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
 3601 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
 3661 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
 3721 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
 3781 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
 3841 gagtccaacc cggtaagaca cgacttatcg ccactggcag ctggtaacag gattagcaga
 3901 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
 3961 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
 4021 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
 4081 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
 4141 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
 4201 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
 4261 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
 4321 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
 4381 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
 4441 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
 4501 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
 4561 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
 4621 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
 4681 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
 4741 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
 4801 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
 4861 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
 4921 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
 4981 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
 5041 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
 5101 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
 5161 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
 5221 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaaatt
 5281 gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
 5341 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
 5401 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
 5461 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
 5521 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
 5581 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
 5641 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
 5701 gccgcgctta atgcgccgct acagggcgcg tcccattcgc ca
//

pET31b载体质粒图谱pdf版和相关资料下载

pET31b载体应用举例

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